JoVE Logo

Sign In

King's College London

43 ARTICLES PUBLISHED IN JoVE

image

Biology

A Novel RFP Reporter to Aid in the Visualization of the Eye Imaginal Disc in Drosophila
Aamna K. Kaul 1, Joseph M. Bateman 1
1Wolfson Centre for Age-Related Diseases, King's College London

We describe a novel red fluorescent protein (RFP) reporter that is expressed specifically in the Drosophila eye. We detail a methodology for dissection of the eye imaginal disc and how this reporter can be used to aid in the dissection and identification of specific cell types in the developing eye.

image

Biology

Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations
Iwan R. Evans 1, Jennifer Zanet 2, Will Wood 1, Brian M. Stramer 2
1Department of Biology and Biochemistry, University of Bath, 2Randall Division of Cell and Molecular Biophysics, King's College London

Drosophila hemocytes disperse over the entirety of the developing embryo. This protocol demonstrates how to mount and image these migrations using embryos with fluorescently labelled hemocytes.

image

Neuroscience

Single Drosophila Ommatidium Dissection and Imaging
Vera Volpi 1, Daniel Mackay 1, Manolis Fanto 1
1MRC Centre for Developmental Neurobiology, King's College London

The limiting factor in the use of the adult Drosophila eye to study neurodegeneration and cell biology is the difficult imaging of intracellular processes. We describe the dissection of single ommatidia to generate a bona-fide primary neuronal cell culture, which can be subject to drug treatment and advanced imaging.

image

Bioengineering

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
Klaus Suhling 1, James A. Levitt 1, Pei- Hua Chung 1, Marina. K. Kuimova 2, Gokhan Yahioglu 3
1Department of Physics, King's College London, 2Department of Chemistry, Imperial College London , 3PhotoBiotics Ltd

Fluorescence Lifetime Imaging (FLIM) has emerged as a key technique to image the environment and interaction of specific proteins and dyes in living cells. FLIM of fluorescent molecular rotors allows mapping of viscosity in living cells.

image

Biology

Electroporation of Craniofacial Mesenchyme
Jacqueline M. Tabler 1, Karen J. Liu 1
1Department of Craniofacial Development, King's College London

Craniofacial cartilages develop in close contact with other tissues and are difficult to manipulate in live animals. We are using electroporation to deliver molecular tools during growth of the craniofacial skeleton while bypassing early embryonic effects. This approach will allow us to efficiently test candidate molecules in vivo.

image

Biology

The Slice Culture Method for Following Development of Tooth Germs In Explant Culture
Sarah A. Alfaqeeh 1,2, Abigail S. Tucker 1
1Department of Craniofacial Development and Stem Cell Biology, and Department of Orthodontics, Dental Institute, Guy's Hospital, UK, King's College London, 2Department of Pediatric Dentistry and Orthodontics, College of Dentistry, King Saud University, Kingdom of Saudi Arabia

Here we detail a method to culture tooth germs in mandible slices using a tissue chopper. This method allows unique access to the tooth during development, providing excellent opportunity for manipulation and lineage tracing, not available using more traditional culture methods.

image

Biology

Rapid Generation of Amyloid from Native Proteins In vitro
Stephanie M Dorta-Estremera 1, Jingjing Li 1, Wei Cao 1
1Department of Immunology, The University of Texas MD Anderson Cancer Center

Proteins can either adopt a native structure or misfold into insoluble amyloid. Conditions that favor the misfolding pathway lead to the formation of different types of amyloid fibrils. The methods described here allow rapid conversion of native proteins into amyloid in vitro.

image

Neuroscience

In vivo Postnatal Electroporation and Time-lapse Imaging of Neuroblast Migration in Mouse Acute Brain Slices
Martina Sonego *1, Ya Zhou *1, Madeleine Julie Oudin 2, Patrick Doherty 1, Giovanna Lalli 1
1Wolfson Centre for Age-Related Diseases, King's College London, 2David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology

Neuroblast migration is a fundamental event in postnatal neurogenesis. We describe a protocol for efficient labeling of neuroblasts by in vivo postnatal electroporation and subsequent visualization of their migration using time-lapse imaging of acute brain slices. We include a description for the quantitative analysis of neuroblast dynamics by video tracking.

image

Neuroscience

Nucleofection of Rodent Neuroblasts to Study Neuroblast Migration In vitro
Katarzyna Falenta *1, Sangeetha Gajendra *2, Martina Sonego 1, Patrick Doherty 1, Giovanna Lalli 1
1Wolfson Centre for Age-Related Diseases, King's College London, 2MRC Centre for Developmental Neurobiology, King's College London

Neuroblast migration is a crucial step in postnatal neurogenesis. The protocol described here can be used to investigate the role of candidate regulators of neuroblast migration by employing DNA/small hairpin RNA (shRNA) nucleofection and a 3D migration assay with neuroblasts isolated from the rodent postnatal rostral migratory stream.

image

Neuroscience

Unilateral Pyramidotomy of the Corticospinal Tract in Rats for Assessment of Neuroplasticity-inducing Therapies
Claudia Kathe 1, Thomas H. Hutson 1, Qin Chen 2, Harold D. Shine 2, Stephen B. McMahon 1, Lawrence D. F. Moon 1
1Neurorestoration, Wolfson Centre for Age-Related Diseases, King's College London, 2Department of Neuroscience, Baylor College of Medicine

The corticospinal tract, one of the major sensorimotor tracts, can be lesioned unilaterally in the rodent brainstem in order to test neuroplasticity-inducing therapies for the central nervous system. This surgical procedure (“pyramidotomy”) and postoperative assessments are described in this protocol.

image

Medicine

From a 2DE-Gel Spot to Protein Function: Lesson Learned From HS1 in Chronic Lymphocytic Leukemia
Benedetta Apollonio 1,2, Maria Teresa Sabrina Bertilaccio 1, Umberto Restuccia 3, Pamela Ranghetti 1, Federica Barbaglio 1, Paolo Ghia 1,4, Federico Caligaris-Cappio 1,4, Cristina Scielzo 1
1Division of Molecular Oncology, IRCCS, San Raffaele Scientific Institute, 2Department of Haemato-Oncology, King's College London, 3IFOM, FIRC Institute of Molecular Oncology, 4Università Vita-Salute San Raffaele

Here we describe a protocol that couples two proteomic techniques, namely 2-dimensional Electrophoresis (2DE) and Mass Spectrometry (MS), to identify differentially expressed/post-translational modified proteins among two or more groups of primary samples. This approach, together with functional experiments, allows the identification and characterization of prognostic markers/therapeutic targets.

image

Developmental Biology

Isolation and Quantitative Immunocytochemical Characterization of Primary Myogenic Cells and Fibroblasts from Human Skeletal Muscle
Chibeza C. Agley 1,2, Anthea M. Rowlerson 1, Cristiana P. Velloso 1, Norman L. Lazarus 1, Stephen D. R. Harridge 1
1Centre of Human and Aerospace Physiological Sciences, King's College London, 2Wellcome Trust-Medical Research Council, Cambridge Stem Cell Institute

The main adherent cell types derived from human muscle are myogenic cells and fibroblasts. Here, cell populations are enriched using magnetic-activated cell sorting based on the CD56 antigen. Subsequent immunolabelling with specific antibodies and use of image analysis techniques allows quantification of cytoplasmic and nuclear characteristics in individual cells.

image

Neuroscience

Investigating the Function of Deep Cortical and Subcortical Structures Using Stereotactic Electroencephalography: Lessons from the Anterior Cingulate Cortex
Robert A. McGovern 1,3, Tarini Ratneswaren 4, Elliot H. Smith 1,3, Jennifer F. Russo 3, Amy C. Jongeling 2,3, Lisa M. Bateman 2,3, Catherine A. Schevon 2,3, Neil A. Feldstein 1,3, Guy M. McKhann, II 1,3, Sameer Sheth 1,3
1Department of Neurosurgery, Columbia University Medical Center, New York Presbyterian Hospital, 2Department of Neurology, Columbia University Medical Center, New York Presbyterian Hospital, 3Columbia University Medical Center, New York Presbyterian Hospital, 4School of Medicine, King's College London

Stereotactic Electroencephalography (SEEG) is an operative technique used in epilepsy surgery to help localize seizure foci. It also affords a unique opportunity to investigate brain function. Here we describe how SEEG can be used to investigate cognitive processes in human subjects.

image

Immunology and Infection

Kupffer Cell Isolation for Nanoparticle Toxicity Testing
Maxime Bourgognon 1, Rebecca Klippstein 1, Khuloud T. Al-Jamal 1
1Institute of Pharmaceutical Science, King's College London

Liver macrophages, named Kupffer cells, are responsible for the capture of circulating nanoparticles. We describe here a method, of high cell purity and yield, for Kupffer cell isolation. The modified LDH assay is used here to measure the toxicity induced by carbon nanotubes in Kupffer cells.

image

Developmental Biology

Ex Vivo Culture of Chick Cerebellar Slices and Spatially Targeted Electroporation of Granule Cell Precursors
Michalina Hanzel 1, Richard J.T. Wingate 1, Thomas Butts 2
1MRC Centre of Developmental Neurobiology, King's College London, 2School of Biological and Chemical Sciences, Queen Mary, University of London

The cerebellar external granule layer is the site of the largest transit amplification in the developing brain. Here, we present a protocol to target genetic modification to this layer at the peak of proliferation using ex vivo electroporation and culture of cerebellar slices from embryonic Day 14 chick embryos.

image

Medicine

A Pipeline for 3D Multimodality Image Integration and Computer-assisted Planning in Epilepsy Surgery
Mark Nowell 1, Roman Rodionov 1, Gergely Zombori 2, Rachel Sparks 2, Michele Rizzi 1, Sebastien Ourselin 2, Anna Miserocchi 3, Andrew McEvoy 3, John Duncan 1
1Department of Clinical and Experimental Epilepsy, UCL Institute of Neurology, 2Center of Medical Imaging and Computing, UCL, 3Department of Neurosurgery, National Hospital for Neurology and Neurosurgery

We describe the steps to use our custom designed software for image integration, visualization and planning in epilepsy surgery.

image

Medicine

The Monoiodoacetate Model of Osteoarthritis Pain in the Mouse
Thomas Pitcher 1, João Sousa-Valente 1, Marzia Malcangio 1
1Wolfson Centre for Age Related Diseases, King's College London

Osteoarthritis (OA), or degenerative joint disease, is a debilitating condition associated with pain that remains only partially controlled by available analgesics. Animal models are being developed to improve our understanding of OA-related pain mechanisms. Here we describe the methodology for the monoiodoacetate model of OA pain in the mouse.

image

Immunology and Infection

Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data
George Ashdown 1, Elvis Pandžić 3, Andrew Cope 2, Paul Wiseman 4, Dylan Owen 1
1Department of Physics and Randall Division of Cell and Molecular Biophysics, King's College London, 2Academic Department of Rheumatology, Centre for Molecular and Cellular Biology of Inflammation, Division of Immunology, Infection and Inflammatory Disease, King's College London, 3ARC Centre for Advanced Molecular Imaging, Australian Centre for NanoMedicine, University of New South Wales Australia, 4Departments of Chemistry and Physic, McGill University

To investigate flow velocities and directionality of filamentous-actin at the T cell immunological synapse, live-cell super-resolution imaging is combined with total internal reflection fluorescence and quantified with spatio-temporal image correlation spectroscopy.

image

Neuroscience

Assessing Primary Neurogenesis in Xenopus Embryos Using Immunostaining
Siwei Zhang *1,2, Jingjing Li *1,3, Robert Lea 1, Enrique Amaya 1
1The Healing Foundation Centre, Faculty of Life Sciences, University of Manchester, 2Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, 3Department of Craniofacial Development and Stem Cell Biology, Dental Institute, King's College London

This article presents a convenient and rapid method for visualizing different neuronal cell populations in the central nervous system of Xenopus embryos using immunofluorescent staining on sections.

image

Developmental Biology

Imaging Cleared Embryonic and Postnatal Hearts at Single-cell Resolution
Wasay M. Shaikh Qureshi 1, Lianjie Miao 1, David Shieh 1, Jingjing Li 1, Yangyang Lu 1, Saiyang Hu 1, Margarida Barroso 1, Joseph Mazurkiewicz 2, Mingfu Wu 1
1Department of Molecular and Cellular Physiology, Albany Medical College, 2Department of Neuroscience and Experimental Therapeutics, Albany Medical College

We describe a protocol to volumetrically image fluorescent protein labeled cells deep inside intact embryonic and postnatal hearts. Utilizing tissue-clearing methods in combination with whole mount staining, single fluorescent protein-labeled cells inside an embryonic or postnatal heart can be imaged clearly and accurately.

image

Behavior

Sit-to-stand-and-walk from 120% Knee Height: A Novel Approach to Assess Dynamic Postural Control Independent of Lead-limb
Gareth D. Jones 1,2, Darren C. James 3, Michael Thacker 1,2, David A. Green 1
1Centre for Human and Aerospace Physiological Sciences (CHAPS), Faculty of Life Sciences and Medicine, King's College London, 2Physiotherapy Department, Guy's & St Thomas' NHS Foundation Trust, London, 3School of Applied Sciences, London South Bank University

Here, we present a novel protocol to measure positional stability at key events during the sit-to-stand-to-walk using the center-of-pressure to the whole-body-center-of-mass distance. This was derived from the force platform and three-dimensional motion-capture technology. The paradigm is reliable and can be utilized for the assessment of neurologically compromised individuals.

image

Bioengineering

Medical-grade Sterilizable Target for Fluid-immersed Fetoscope Optical Distortion Calibration
Daniil I. Nikitichev *1, Dzhoshkun I. Shakir *1, François Chadebecq 1, Marcel Tella 1, Jan Deprest 1,2, Danail Stoyanov 3, Sébastien Ourselin 1, Tom Vercauteren 1
1Translational Imaging Group, CMIC, University College London, 2Department of Obstetrics and Gynecology, University Hospitals Leuven, 3Surgical Robot Vision Group, CMIC, University College London

This article describes the design and development of a sterilizable custom camera optical distortion calibration target for the peri-operative, fluid-immersed calibration of endoscopes during endoscopic interventions.

image

JoVE Journal

Glycoproteomics of the Extracellular Matrix: A Method for Intact Glycopeptide Analysis Using Mass Spectrometry
Javier Barallobre-Barreiro 1, Ferheen Baig 1, Marika Fava 1, Xiaoke Yin 1, Manuel Mayr 1
1King's British Heart Foundation Centre, King's College London

This paper describes a methodology to prepare cardiovascular tissue samples for MS analysis that allows for (1) the analysis of ECM protein composition, (2) the identification of glycosylation sites, and (3) the compositional characterization of glycan forms. This methodology can be applied, with minor modifications, to the study of the ECM in other tissues.

image

Developmental Biology

Horizontal Whole Mount: A Novel Processing and Imaging Protocol for Thick, Three-dimensional Tissue Cross-sections of Skin
Lucia Salz 1,2, Ryan R. Driskell 1,2
1Centre for Stem Cells and Regenerative Medicine, King's College London, 2School of Molecular Biosciences, Washington State University

This work presents a novel processing and imaging protocol for thick, three-dimensional tissue cross-section analysis that enables the full exploitation of confocal imaging modalities. This protocol preserves antigenicity and represents a robust system to analyze skin histology and potentially other tissue types.

image

Medicine

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells
Simone S. E. Nielsen 1, Piotr Siupka 1, Ana Georgian 2, Jane E. Preston 2, Andrea E. Tóth 1, Siti R. Yusof 2,3, N. Joan Abbott 2, Morten S. Nielsen 1
1Lundbeck Foundation Research Initiative on Brain Barriers and Drug Delivery, Department of Biomedicine, Aarhus University, 2Institute of Pharmaceutical Science, King's College London, 3HICoE Centre for Drug Research, Universiti Sains Malaysia

The aim of the protocol is to present an optimized procedure for the establishment of an in vitro blood-brain barrier (BBB) model based on primary porcine brain endothelial cells (pBECs). The model shows high reproducibility, high tightness, and is suitable for studies of transport and intracellular trafficking in drug discovery.

image

JoVE Journal

Quantification of Information Encoded by Gene Expression Levels During Lifespan Modulation Under Broad-range Dietary Restriction in C. elegans
Dhaval S Patel *1, Giovanni Diana *1, Eugeni V. Entchev 1, Mei Zhan 2,3,4, Hang Lu 2,3,4, QueeLim Ch'ng 1
1Centre for Developmental Neurobiology, King's College London, 2Interdisciplinary Bioengineering Graduate Program, Georgia Institute of Technology, 3Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 4School of Chemical & Biomolecular Engineering, Georgia Institute of Technology

Here, we present a framework to relate broad-range dietary restriction to gene expression and lifespan. We describe protocols for broad-range dietary restriction and for quantitative imaging of gene expression under this paradigm. We further outline computational analyses to reveal underlying information processing features of the genetic circuits involved in food-sensing.

image

JoVE Journal

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
Caroline Haupt *1, Tommy Hofmann *1, Sabine Wittig *1, Susann Kostmann 1, Argyris Politis 2, Carla Schmidt 1
1Interdisciplinary research center HALOmem, Martin Luther University Halle-Wittenberg, 2Department of Chemistry, Kings College London

The architecture of protein complexes is essential for their function. Combining various mass spectrometric techniques proved powerful to study their assembly. We provide protocols for chemical cross-linking and native mass spectrometry and show how these complementary techniques help to elucidate the architecture of multi-subunit protein assemblies.

image

Cancer Research

Radionuclide-fluorescence Reporter Gene Imaging to Track Tumor Progression in Rodent Tumor Models
Alessia Volpe 1, Francis Man 1, Lindsay Lim 1, Alex Khoshnevisan 1, Julia Blower 1, Philip J. Blower 1, Gilbert O. Fruhwirth 1
1Department of Imaging Chemistry and Biology, School of Biomedical Engineering and Imaging Sciences, King's College London

We describe a protocol for preclinical in vivo tracking of cancer metastasis. It is based on a radionuclide-fluorescence reporter combining the sodium iodide symporter, detected by non-invasive [18F]tetrafluoroborate-PET, and a fluorescent protein for streamlined ex vivo confirmation. The method is applicable for preclinical in vivo cell tracking beyond tumor biology.

image

Engineering

Three-Dimensional Ultrasonic Needle Tip Tracking with a Fiber-Optic Ultrasound Receiver
Wenfeng Xia 1,2, Simeon J. West 3, Malcolm C. Finlay 2,4, Rosalind Pratt 5,6, Sunish Mathews 1,2, Jean-Martial Mari 7, Sebastien Ourselin 1,2,6, Anna L. David 1,5,8,9, Adrien E. Desjardins 1,2
1Wellcome/EPSRC Centre for Interventional and Surgical Sciences, University College London, 2Department of Medical Physics and Biomedical Engineering, University College London, 3Department of Anaesthesia, University College Hospital, 4St Bartholomew's Hospital and Queen Mary University of London, 5Institute for Women's Health, University College London, 6Centre for Medical Imaging Computing, University College London, 7GePaSud, University of French Polynesia, 8Department of Development and Regeneration, KU Leuven (Katholieke Universiteit), 9NIHR University College London Hospitals Biomedical Research Centre

Accurate and efficient visualization of invasive medical devices is extremely important in many ultrasound-guided minimally invasive procedures. Here, a method for localizing the spatial position of a needle tip relative to the ultrasound imaging probe is presented.

image

Research

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry
Zainab Ahdash 1, Andy M. Lau 1, Chloe Martens 1, Argyris Politis 1
1Department of Chemistry, King's College London

Mass spectrometry (MS) has emerged as an important tool for the investigation of structure and dynamics of macromolecular assemblies. Here, we integrate MS-based approaches to interrogate protein complex formation and ligand binding.

image

Immunology and Infection

Quantitative Polymerase Chain Reaction-based Analyses of Murine Intestinal Microbiota After Oral Antibiotic Treatment
Rebeca Jimeno 1,2, Phillip M. Brailey 1,2, Patricia Barral 1,2
1The Peter Gorer Department of Immunobiology, King's College London, 2The Francis Crick Institute

Here we provide detailed protocols for the oral administration of antibiotics to mice, collection of fecal samples, DNA extraction and quantification of fecal bacteria by qPCR.

image

Genetics

Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
Darja Pollpeter 1, Andrew Sobala 1, Michael H. Malim 1
1Department of Infectious Diseases, School of Immunology & Microbial Sciences, King's College London

Here we present a deep sequencing approach that provides an unbiased determination of nascent 3'-termini as well as mutational profiles of single-stranded DNA molecules. The main application is the characterization of nascent retroviral complementary DNAs (cDNAs), the intermediates generated during the process of retroviral reverse transcription.

image

Engineering

Design and Implementation of a Bespoke Robotic Manipulator for Extra-corporeal Ultrasound
Shuangyi Wang 1, James Housden 1, Yohan Noh 1, Anisha Singh 2, Junghwan Back 3, Lukas Lindenroth 3, Hongbin Liu 3, Joseph Hajnal 1, Kaspar Althoefer 4, Davinder Singh 2, Kawal Rhode 1
1School of Biomedical Engineering & Imaging Sciences, King's College London, 2Xtronics Ltd, 3Department of Informatics, King's College London, 4Faculty of Science & Engineering, Queen Mary University of London

This paper introduces the design and implementation of a bespoke robotic manipulator for extra-corporeal ultrasound examination. The system has five degrees of freedom with lightweight joints made by 3D printing and a mechanical clutch for safety management.

image

Cancer Research

Preparation of Exosomes for siRNA Delivery to Cancer Cells
Farid N. Faruqu *1, Lizhou Xu *1, Khuloud T. Al-Jamal 1
1Institute of Pharmaceutical Science, King's College London

An exosome is a new generation of drug delivery carriers. We established an exosome isolation protocol with high yield and purity for siRNA delivery. We also encapsulated fluorescently labelled non-specific siRNA into exosomes and investigated the cellular uptake of siRNA-loaded exosomes in cancer cells.

image

Neuroscience

Transplantation of Chemogenetically Engineered Cortical Interneuron Progenitors into Early Postnatal Mouse Brains
Myrto Denaxa 1,2, Guilherme Neves 3, Juan Burrone 3, Vassilis Pachnis 1
1Nervous System Development and Homeostasis Laboratory, The Francis Crick Institute, 2Neuroscience Centre, Biomedical Sciences Research Centre "Al. Fleming", 3Centre for Developmental Neurobiology, King's College London

Here we present a protocol, designed to use chemogenetic tools to manipulate the activity of cortical interneuron progenitors transplanted into the cortex of early postnatal mice.

image

Neuroscience

Meta-analysis of Voxel-Based Neuroimaging Studies using Seed-based d Mapping with Permutation of Subject Images (SDM-PSI)
Anton Albajes-Eizagirre 1,2, Aleix Solanes 1,2, Miquel Angel Fullana 2,3, John P. A. Ioannidis 4, Paolo Fusar-Poli 5,6,7, Carla Torrent 1,2,3,8, Brisa Solé 1,2,3,8, Caterina Mar Bonnín 1,2,3,8, Eduard Vieta 1,2,3,8, David Mataix-Cols 9, Joaquim Radua 1,2,5,9
1Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 2Mental Health Research Networking Center (CIBERSAM), 3Institute of Neurosciences, Hospital Clinic de Barcelona, 4Departments of Medicine, of Health Research and Policy, and of Biomedical Data Science, Stanford University School of Medicine, and Department of Statistics, Stanford University School of Humanities and Sciences, 5Department of Psychosis Studies, Institute of Psychiatry, Psychology and Neuroscience, King's College London, 6OASIS Service, South London and Maudsley NHS Foundation Trust, 7Department of Nervous System and Behavioral Sciences, University of Pavia, 8University of Barcelona, 9Centre for Psychiatric Research and Education, Department of Clinical Neuroscience, Karolinska Institutet

We detail how to conduct a meta-analysis of voxel-based neuroimaging studies using Seed-based d Mapping with Permutation of Subject Images (SDM-PSI).

image

Biology

Assessing Mineral Availability in Fish Feeds using Complementary Methods Demonstrated with the Example of Zinc in Atlantic Salmon
Marta S. Silva 1,2, Thea Stewart 4, Heidi Amlund 1,3, Jens J. Sloth 1,3, Pedro Araujo 1, Erik-Jan Lock 1, Christer Hogstrand 4, Robin Ørnsrud 1, Rune Waagbø 1,2, Antony Jesu Prabhu 1
1Institute of Marine Research, 2Department of Biological Sciences, University of Bergen, 3National Food Institute, Technical University of Denmark, 4Metal metabolism group, Nutritional Sciences Division, King's College London

This article explains in detail a systematic approach to assess micro-mineral availability in Atlantic salmon. The methodology includes tools and models with increasing biological complexity: (1) chemical speciation analysis, (2) in vitro solubility, (3) uptake studies in cell lines, and (4) in vivo fish studies. 

image

Developmental Biology

Dissection, Culture and Analysis of Primary Cranial Neural Crest Cells from Mouse for the Study of Neural Crest Cell Delamination and Migration
Sandra Guadalupe Gonzalez Malagon *1,2, Lisa Dobson *1,3, Anna M Lopez Muñoz 1, Marcus Dawson 1, William Barrell 1,3, Petros Marangos 2,4, Matthias Krause 3, Karen J Liu 1
1Centre for Craniofacial and Regenerative Biology, King's College London, 2Institute of Molecular Biology and Biotechnology, FORTH, Department of Biomedical Research, University of Ioannina, 3Randall Centre of Cell & Molecular Biophysics, King's College London, 4Department of Biological Applications and Technology, University of Ioannina

This protocol describes the dissection and culture of cranial neural crest cells from mouse models, primarily for the study of cell migration. We describe the live imaging techniques used and the analysis of speed and cell shape changes.

image

Biology

Quantification of Proliferative and Dead Cells in Enteroids
Hua-Shan Li *1, Shao-Fang Xu *1, Jian-Ying Sheng *1, Zhi-Hui Jiang 1, Jing Wang 1, Ning Ding 1, Tao Wang 1, Matthew A. Odenwald 2, Jerrold R. Turner 2,3, Wei-Qi He 1, Hong Xu 1, Juan-Min Zha 1
1Jiangsu Key Laboratory of Neuropsychiatric Diseases and Cambridge-Suda (CAM-SU) Genomic Resource Center, Medical College of Soochow University, Department of Oncology, The First Affiliated Hospital of Soochow University, 2Department of Pathology, University of Chicago, 3Department of Pathology, Brigham and Women's Hospital–Harvard Medical School

The presented protocol uses flow cytometry to quantify the number of proliferating and dead cells in cultured mouse enteroids. This method is helpful to evaluate the effects of drug treatment on organoid proliferation and survival.

image

Behavior

Implementation of a Real-Time Psychosis Risk Detection and Alerting System Based on Electronic Health Records using CogStack
Tao Wang 1, Dominic Oliver 2, Yamiko Msosa 1, Craig Colling 3, Giulia Spada 2, Łukasz Roguski 4, Amos Folarin 1, Robert Stewart 3,5, Angus Roberts 1,3, Richard J. B. Dobson 1,3,4,6, Paolo Fusar-Poli 2,3,7,8
1Department of Biostatistics and Health Informatics, Institute of Psychiatry, Psychology and Neuroscience, King's College London, 2Early Psychosis: Interventions and Clinical-detection (EPIC) lab, Department of Psychosis Studies, Institute of Psychiatry, Psychology & Neuroscience, King's College London, 3National Institute for Health Research, Maudsley Biomedical Research Centre, South London and Maudsley National Health Service (NHS) Foundation Trust, 4Institute of Health Informatics, University College London, 5Department of Psychological Medicine, Institute of Psychiatry, Psychology and Neuroscience, King's College London, 6Health Data Research UK London, University College London, 7OASIS service, South London and Maudsley National Health Service (NHS) Foundation Trust, 8Department of Brain and Behavioral Sciences, University of Pavia

We demonstrate how to deploy a real-time psychosis risk calculation and alerting system based on CogStack, an information retrieval and extraction platform for electronic health records.

image

Medicine

Applying a Three-dimensional Uniaxial Mechanical Stimulation Bioreactor System to Induce Tenogenic Differentiation of Tendon-Derived Stem Cells
Ziming Chen *1, Peilin Chen *1, Rui Ruan *1, Lianzhi Chen 1, Jun Yuan 1, David Wood 1, Tao Wang 1, Ming Hao Zheng 1
1Centre of Orthopaedic Translational Research, Medical School, University of Western Australia

A three-dimensional uniaxial mechanical stimulation bioreactor system is an ideal bioreactor for tenogenic-specific differentiation of tendon-derived stem cells and neo-tendon formation.

image

Bioengineering

Patient-Specific Polyvinyl Alcohol Phantom Fabrication with Ultrasound and X-Ray Contrast for Brain Tumor Surgery Planning
Eleanor C. Mackle *1,2, Jonathan Shapey *1,2,3,4, Efthymios Maneas 1,2, Shakeel R. Saeed 3,5,6, Robert Bradford 3, Sebastien Ourselin 4, Tom Vercauteren 4, Adrien E. Desjardins 1,2
1Wellcome / EPSRC Centre for Interventional and Surgical Sciences, University College London, 2Department of Medical Physics and Biomedical Engineering, University College London, 3Department of Neurosurgery, National Hospital for Neurology and Neurosurgery, 4School of Biomedical Engineering & Imaging Sciences, King's College London, 5The Ear Institute, University College London, 6The Royal National Throat, Nose and Ear Hospital, London

This protocol describes the fabrication of a patient specific skull, brain and tumor phantom. It uses 3D printing to create molds, and polyvinyl alcohol (PVA-c) is used as the tissue mimicking material.

image

Biology

Novel In Vivo Micro-Computed Tomography Imaging Techniques for Assessing the Progression of Non-Alcoholic Fatty Liver Disease
Anna Hadjihambi *1,2, Rallia-Iliana Velliou *3, Panagiotis Tsialios 4, Aigli-Ioanna Legaki 3, Antonios Chatzigeorgiou *3, Maritina G. Rouchota *4
1The Roger Williams Institute of Hepatology London, Foundation for Liver Research, 2Faculty of Life Sciences and Medicine, King's College London, 3Department of Physiology, Medical School, National and Kapodistrian University of Athens, 4BIOEMTECH, Lefkippos Attica Technology Park NCSR "Demokritos"

Using a diet-induced non-alcoholic fatty liver disease (NAFLD) mouse model, we describe the use of novel in vivo micro-computed tomography imaging techniques as a non-invasive method to assess the progression stages of NAFLD, focusing predominantly on the hepatic vascular network due to its significant involvement in NAFLD-related hepatic dysregulation.

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved